Ureylene phenylene anionic naphthalenesulfonic acids

ABSTRACT

Novel ureylenebis[substituted-phenylenecarbonyl(and sulfonyl)imino-substituted-phenylenesulfonylimino-naphthalenetrisulfonic acid hexaalkali metal salts], useful as inhibitors of the complement system of warm-blooded animals, the amino-substituted phenylenecarbonyl (and sulfonyl)imino-substituted-phenylenesulfonylimino-naphthalenetrisulfonic acid, trialkali metal salts, which are new intermediates for the preparation of the active ureylenes, and the process for their preparation.

This is a division of application Ser. No. 923,742, filed July 11, 1978.

BACKGROUND OF THE INVENTION

The term "complement" refers to a complex group of proteins in bodyfluids that, working together with antibodies or other factors, play animportant role as mediators of immune, allergic, immunochemical and/orimmunopathological reactions. The reactions in which complementparticipates take place in blood serum or in other body fluids, andhence are considered to be humoral reactions.

With regard to human blood, there are at present more than 11 proteinsin the complement system. These complement proteins are designated bythe letter C and by number: C1, C2, C3 and so on up to C9. Thecomplement protein C1 is actually an assembly of subunits designatedC1q, C1r and C1s. The numbers assigned to the complement proteinsreflect the sequence in which they become active, with the exception ofcomplement protein C4, which reacts after C1 and before C2. Thenumerical assignments for the proteins in the complement system weremade before the reaction sequence was fully understood. A more detaileddiscussion of the complement system and its role in the body processescan be found in, for example, Bull. World Health Org., 39, 935-938(1968); Ann. Rev. Medicine, 19, 1-24 (1968); The John Hopkins Med. J.,128, 57-74 (1971); Harvey Lectures, 66, 75-104 (1972); The New EnglandJournal of Medicine, 287, 452-454; 489-495; 545-549; 592-596; 642-646(1972); Scientific American, 229, (No. 5), 54-66 (1973); FederationProceedings, 32, 134-137 (1973); Medical World News, October 11, 1974,pp. 53-66; J. Allergy Clin. Immunol., 53, 298-302 (1974); Cold SpringHarbor Conf. Cell Prolifieration 2/Proteases Biol. Control/229-241(1975); Ann. Review of Biochemistry, 44, 697 (1975); Complement inClinical Medicine, Disease-a-Month, (1975); Complement, Scope, December1975; Annals of Internal Medicine, 84, 580-593 (1976); "Complement:Mechanisms and Functions", Prentice-Hall, Englewood Cliffs, N.J. (1976);Essays Med. Biochem., 2, 1-35 (1976); Hospital Practice, 12, 33-43(1977); Perturbation of Complement in Disease, Chap. 15 in BiologicalAmplification Systems in Immunology (Ed. Day and Good), Plenum, New Yorkand London (1977); Am. J. Clin. Pathology, 68, 647-659 (1977).

The complement system can be considered to consist of three sub-systems:(1) a recognition unit (C1q) which enables it to combine with antibodymolecules that have detected a foreign invader; (2) an activation unit(C1r, C1s, C2, C4, C3) which prepares a site on the neighboringmembrane; and (3) an attack unit (C5, C6, C7, C8 and C9) which creates a"hole" in the membrane. The membrane attack unit is non-specific; itdestroys invaders only because it is generated in their neighborhood. Inorder to minimize damage to the host's own cells, its activity must belimited in time. This limitation is accomplished partly by thespontaneous decay of activated complement and partly by interference byinhibitors and destructive enzymes. The control of complement, however,is not perfect, and there are times when damage is done to the host'scells. Immunity is, therefore, a double-edged sword.

Activation of the complement system also accelerates blood clotting.This action comes about by way of the complement-mediated release of aclotting factor from platelets. The biologically active complementfragments and complexes can become involved in reactions that damage thehost's cells, and these pathogenic reactions can result in thedevelopment of immune-complex diseases. For example, in some forms ofnephritis, complement damages the basal membrane of the kidney,resulting in the escape of protein from the blood into the urine. Thedisease disseminated lupus erythematosus belongs in this category; itssymptoms include nephritis, visceral lesions and skin eruptions. Thetreatment of diphtheria or tetanus with the injection of large amountsof antitoxin sometimes results in serum sickness, an immune-complexdisease. Rheumatoid arthritis also involves immune complexes. Likedisseminated lupus erythematosus, it is an autoimmune disease in whichthe disease symptoms are caused by pathological effects of the immunesystem in the host's tissues. In summary, the complement system has beenshown to be involved with inflammation, coagulation, fibrinolysis,antibody-antigen reactions and other metabolic processes.

In the presence of antibody-antigen complexes the complement proteinsare involved in a series of reactions which may lead to irreversiblemembrane damage if they occur in the vicinity of biological membranes.Thus, while complement constitutes a part of the body's defensemechanism against infection it also results in inflammation and tissuedamage in the immunopathological process. The nature of certain of thecomplement proteins, suggestions regarding the mode of complementbinding to biological membranes and the manner in which complementeffects membrane damage are discussed in Annual Review in Biochemistry,38, 389 (1969); Journal of Immunology, 119, 1-8, 1195, 1358-1364, 1482(1977).

A variety of substances have been disclosed as inhibiting the complementsystem, i.e., as complement inhibitors. For example, the compounds3,3'-ureylenebis[6-(2-amino-8-hydroxy-6-sulfo-1-naphthylazo)benzenesulfonicacid], tetrasodium salt (chlorazol fast pink), heparin and a sulphateddextran have been reported to have an anticomplementary effect, BritishJournal of Experimental Pathology, 33, 327-339 (1952). German Pat. No.2,254,893 or South African Pat. No. 727,923 discloses certain1-(diphenylmethyl)-4-(3-phenylallyl)piperazines useful as complementinhibitors. Other chemical compounds having complement inhibitingactivity are disclosed in, for example, Journal of Medicinal Chemistry,12, 415-419; 902-905; 1049-1052; 1053-1056 (1969); Canadian Journal ofBiochemistry, 47, 547-552 (1969); The Journal of Immunology, 104,279-288 (1970); The Journal of Immunology, 106, 241-245 (1971); TheJournal of Immunology, 111, 1061-1066 (1973); Biochim. Biophys. Acta,317, 539-548 (1973); Life Sciences, 13, 351-362 (1973); Journal ofImmunology, 113, 584 (1974); Immunology, 26, 819-829 (1974); Journal ofMedicinal Chemistry, 17, 1160-1167 (1974); Biochim. Biophys. Res. Comm.,67, 225-263 (1975); Ann. N.Y. Acad. Sci., 256, 441-450 (1975); Journalof Medicinal Chemistry, 19, 634-639, 1079 (1976); Journal of Immunology,118, 466 (1977); Arch. Int. Pharmacodyn., 226, 281-285 (1977); Biochem.Pharmacol. 26, 325-329 (1977); Journal Pharm. Sci., 66, 1367-1377(1977); Chem. Pharm. Bull., 25, 1202-1208 (1977); Biochim. Biophys.Acta, 484, 417-422 (1977) and Journal Clin. Microbiology, 5, 278-284(1977).

It has been reported that the known complement inhibitorsepsilon-aminocaproic acid and tranexamic acid have been used withsuccess in the treatment of hereditary angioneurotic edema, a diseasestate resulting from an inherited deficiency or lack of function of theserum inhibitor of the activated first component of complement (C1inhibitor). The New England Journal of Medicine, 286, 808-812 (1972),287, 452-454 (1972); Ann. Intern. Med., 84, 580-593 (1976); J. Allergyand Clin. Immunology, 60, 38-40 (1977).

It has also been reported that the drug pentosan-polysulfoester has ananticomplementary activity on human serum, both in vitro and in vivo, asjudged by the reduction in total hemolytic complement activity;Pathologie Biologie, 25, 33-36, 25 (2), 105-108, 25 (3), 179-184 (1977).

It is known that the compound Suramin is moderately active as acomplement inhibitor, and possesses the structure: ##STR1##

It now has been discovered that certain modifications of this structureprovide compounds with enhanced inhibitory activity. This invention isbased on such modifications.

The following publications, pertaining to the chemistry of Suramin, arerelated to the preparation of the novel compounds of this invention:

Bayer & Co., D.R.P. 278,122, June 22, 1913 [C.A. 9, 1096(1915)]

Bayer & Co., D.R.P. 288,272, Jan. 23, 1914 [C.A. 10, 2279(1916)]

Bayer & Co., D.R.P. 288,273, Feb. 21, 1914 [C.A. 10, 2279(1916)]

Frdl. 12, 185-186, 191-195 (1914-1916)

Danish Pat. No. 20,743 (1915)

Austrian Pat. No. 72,298 (1916)

Austrian Pat. No. 72,303 (1916)

U.S. Pat. No. 1,218,654 (1917)

U.S. Pat. No. 1,218,655 (1917)

Austrian Pat. No. 73,381 (1917)

U.S. Pat. No. 1,308,071 (1919)

E. Fourneau, J. Trefouel, Mme. J. Trefouel and J. Vallee, Acad. Sci.Comp. Rend., 178, 675-676 (1924)

E. Fourneau, F. Trefouel and J. Vallee, Ann. de L'Institut Pasteur, 38(2), 81-114 (1924)

B. Heymann, Zeitschrift Ang. Chem., 37, 585-589 (1924)

British Pat. No. 224,849 (1925)

U.S. Pat. No. 1,606,624 (1926)

J. E. R. McDonagh, Brit. Med. J., 693-696 (1926) [Chem. Zentralblatt,1769-1770 (1926 II)]

W. Roehl, Arch. Schiff. Trop. Hyg., 30 (1), 103-111 (1926)

Poulenc Freres, D.R.P. 427,857, April 20, 1926 [Frdl. 15,1434-1436(1928)]

I. E. Balaban and H. King, J. Chem. Soc., 3068-3097 (1927)

H. Bauer and J. Becker, Arb. Staatsinst. Exptl. Therap., 16 pp. (1928)

U.S. Pat. No. 1,968,820 (1934)

O. Yu. Magidson, O. S. Madaeva and M. V. Rubtzov, Khim. Farm. Prom., 2,89-94 (1935) [C.A., 30, 4492 (1936)]

U.S. Pat. No. 2,126,180 (1938)

P. Pratsi and L. Raffa, Farmaco Sci e Tec (Pavia), 1, 21-34 (1946)

A. Spinks, Biochem. J., 42, 109-116 (1948)

E. D. Wills and A. Wormall, Biochem. J., 47, 158-170 (1950)

German Pat. No. 890,952 (1953) [C. A. 52, 14693 (1958)]

A. Adams, J. N. Ashley and H. Bader, J. Chem. Soc., 3739-3744 (1956) [C.A. 51, 4375i]

Publications related to the biological use of Suramin compounds for thepurpose of inhibiting the complement system, including humans, asdetermined by the in vivo and in vitro testing of the blood serum ofwarm-blooded animals are:

B. Stuber and K. Lang, Arch. Exptl. Path. Pharmacol., 154, 41-49 (1930)[C. A. 25, 3067(1931)]

F. Klopstock, Zeitschrift fur Immunitatsforschung und experimentalleTherapie, 75, 348-354 (1932)

H. J. Schmid, Schweiz. Med. Woch., 96, 1267-1269 (1966)

K. Lauenstein, Bayer-Symposium I, 25-30 (1969)

J. S. C. Fong and R. A. Good, Clin. Exp. Immunol., 10, 127-138 (1972)

V. Eisen and C. Loveday, Br. J. Pharmac., 49, 678-687 (1973)

D. Brackertz and F. Kueppers, Allergol. Et Immunopath., 11, 163-168(1974)

E. Raepple, H-U. Hill and M. Loos, Immunochemistry, 13 (3), 251-255(1976)

SUMMARY OF THE INVENTION

This invention is concerned withureylenebis[substituted-phenylenecarbonyl(andsulfonyl)imino-substituted-phenylenesulfonylimino-naphthalenetrisulfonicacids] and all pharmaceutically acceptable salts thereof, havingcomplement inhibiting activity, which are new compounds of the generalformulae: ##STR2## wherein R, R₁, R₂ and R₃ are selected from the groupconsisting of hydrogen and methyl; and A is a pharmaceuticallyacceptable salt cation.

This invention is also concerned with compounds of the formulae:##STR3## wherein R, R₁, R₂ and R₃ are selected from the group consistingof hydrogen and methyl; and A is a pharmaceutically acceptable saltcation; said compounds being useful as intermediates for the preparationof the complement inhibiting compounds described above. Some of theintermediate compounds also possess complement inhibiting activity.

DESCRIPTION OF THE INVENTION

The novel intermediate amine compounds of the invention are prepared byreacting the appropriate 8-amino-1,3,5(and 1,3,6)-naphthalenetrisulfonicacid trialkali metal salt with a nitrobenzenesulfonyl chloride such asm-nitrobenzenesulfonyl chloride, 3-nitro-p-toluenesulfonyl chloride and2-methyl-5-nitrobenzenesulfonyl chloride, for 1.5-36 hours in an aqueoussolution made alkaline with alkali metal hydroxide, anhydrous alkalimetal carbonate or alkali metal acetate trihydrate. Afterneutralization, the solution is diluted with absolute ethanol to providethe corresponding nitro-substituted-phenylenesulfonylimino-1,3,5(and1,3,6)-naphthalenetrisulfonic acid, trialkali metal salt.

Hydrogenation of the preceding nitro trialkali metal salts using 10%palladium-carbon catalyst, filtration, concentration and treatment withabsolute ethanol provides the correspondingamino-substituted-phenylenesulfonylimino naphthalenetrisulfonic acid,trialkali metal salt compounds.

The amino compounds above, dissolved in aqueous media and made alkalinewith either alkali metal hydroxide or anhydrous alkali metal carbonateare reacted once more with the above listed nitrobenzenesulfonylchloride or a nitrobenzoyl chloride such as m-nitrobenzoyl chloride or3-nitro-p-toluoyl chloride for 1.5-36 hours. After neutralization, thesolution is diluted with absolute ethanol to provide the correspondingnitro-substituted-phenylenecarbonyl (andsulfonyl)imino-substituted-phenylenesulfonylimino-naphthalenetrisulfonicacid, trialkali metal salt.

The novel intermediate amine compounds of the invention are thenobtained by hydrogenation of the above nitro compounds using 10%palladium-carbon catalyst in water as previously described, filtrationand evaporation of the filtrate produces a residue which is dissolved inwater and precipitated with absolute ethanol to provide the desiredproduct.

The novel ureylene compounds of the invention, which are activecomplement inhibitors, are then provided by treatment of the aboveintermediate amine compounds with phosgene in aqueous media madealkaline with alkali metal carbonate or pyridine, neutralization, andprecipitation from aqueous solution with alcohol.

This invention is concerned with a method of inhibiting the complementsystem in a body fluid, such as blood serum, which comprises subjectingbody fluid complement to the action of an effective complementinhibiting amount of a compound encompassed within the formulaehereinabove. The method of use aspect of this invention is alsoconcerned with a method of inhibiting the complement system in awarm-blooded animal which comprises administering to said animal aneffective complement inhibiting amount of a compound encompassed withinthe formulae hereinabove. Body fluid can include blood, plasma, serum,synovial fluid, cerebrospinal fluid, or pathological accumulations offluid such as pleural effusion, etc.

Compounds of the present invention find utility as complement inhibitorsin body fluids and as such may be used to ameliorate or prevent thosepathological reactions requiring the function of complement and in thetherapeutic treatment of warm-blooded animals having immunologicdiseases such as rheumatoid arthritis, systemic lupus erythematosus,certain kinds of glomerulonephritis, certain kinds of auto-allergichemolytic anemia, certain kinds of platelet disorders and certain kindsof vasculitis. The compounds herein may also be used in the therapeutictreatment of warm-blooded animals having non-immunologic diseases suchas paroxysmal nocturnal hemoglobinuria, hereditary angioneurotic edema(treated with Suramin, etc.) and inflammatory states induced by theaction of bacterial or lysosomal enzymes on the appropriate complementcomponents as for example, inflammation following coronary occlusion.They may also be useful in the treatment of transplant rejection and asblood culture or transport mediums.

The compounds of the present invention may be administered internally,e.g., orally, or parenterally, e.g., intra-articularly, to awarm-blooded animal to inhibit complement in the body fluid of theanimal, such inhibition being useful in the amelioration or preventionof those reactions dependent upon the function of complement, such asinflammatory process and cell membrane damage induced byantigen-antibody complexes. A range of doses may be employed dependingon the mode of administration, the condition being treated and theparticular compound being used. For example, for intravenous orsubcutaneous use from about 5 to about 50 mg/kg/day, or every six hoursfor more rapidly excreted salts, may be used. For intra-articular usefor large joints such as the knee, from about 2 to about 20 mg/joint perweek may be used, with proportionally smaller doses for smaller joints.The dosage range is to be adjusted to provide optimum therapeuticresponse in the warm-blooded animal being treated. In general, theamount of compound administered can vary over a wide range to providefrom about 5 mg/kg to about 100 mg/kg of body weight of animal per day.The usual daily dosage for a 70 kg subject may vary from about 350 mg toabout 3.5 g. Unit doses of the acid or salt can contain from about 0.5mg to about 500 mg.

While in general the sodium salts of the acids of the invention aresuitable for parenteral use, other sadts may also be prepared, such asthose of primary amines, e.g., ethylamine; secondary amines, e.g.,diethylamine or diethanol amine; tertiary amines, e.g., pyridine ortriethylamine or 2-dimethylaminomethyldibenzofuran; aliphatic diamines,e.g., decamethylenediamine; and aromatic diamines, can be prepared. Someof these are soluble in water, others are soluble in saline solution,and still others are insoluble and can be used for purposes of preparingsuspensions for injection. Furthermore, as well as the sodium salt,those of the alkali metals, such as potassium and lithium; of ammonia;and of the alkaline earth metals, such as calcium or magnesium, may beemployed. It will be apparent, therefore, that these salts embrace, ingeneral, derivatives of salt-forming cations.

The compounds of the present invention may also be administeredtopically in the form of ointments, creams, lotions and the like,suitable for the treatment of complement dependent dermatologicaldisorders.

Moreover, the compounds of the present invention may be administered inthe form of dental pastes, ointments, buccal tablets and othercompositions suitable for application periodontally for the treatment ofperiodontitis and related diseases of the oral cavity.

In therapeutic use, the compounds of this invention may be administeredin the form of conventional pharmaceutical compositions. Suchcompositions may be formulated so as to be suitable for oral orparenteral administration. The active ingredient may be combined inadmixture with a pharmaceutically acceptable carrier, which carrier maytake a wide variety of forms depending on the form of preparationdesired for administration, i.e., oral or parenteral. The compounds canbe used in compositions such as tablets. Here, the principal activeingredient is mixed with conventional tabletting ingredients such ascorn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesiumstearate, dicalcium phosphate, gums, or similar materials as non-toxicpharmaceutically acceptable diluents or carriers. The tablets or pillsof the novel compositions can be laminated or otherwise compounded toprovide a dosage form affording the advantage of prolonged or delayedaction or predetermined successive action of the enclosed medication.For example, the tablet or pill can comprise an inner dosage and anouter dosage component, the latter being in the form of an envelope overthe former. The two components can be separated by an enteric layerwhich serves to resist disintegration in the stomach and permits theinner component to pass intact into the duodenum or to be delayed inrelease. A variety of materials can be used for such enteric layers orcoatings, such materials including a number of polymeric acids ormixtures of polymeric acids with such materials as shellac, shellac andcetyl alcohol, cellulose acetate and the like. A particularlyadvantageous enteric coating comprises a styrene maleic acid copolymertogether with known materials contributing to the enteric properties ofthe coating. The tablet or pill may be colored through the use of anappropriate non-toxic dye, so as to provide a pleasing appearance.

The liquid forms in which the novel compositions of the presentinvention may be incorporated for administration include suitableflavored emulsions with edible oils, such as, cottonseed oil, sesameoil, coconut oil, peanut oil, and the like, as well as elixirs andsimilar pharmaceutical vehicles. Sterile suspensions or solutions can beprepared for parenteral use. Isotonic preparations containing suitablepreservatives are also desirable for injection use.

The term dosage form, as described herein, refers to physically discreteunits suitable as unitary dosage for warm-blooded animal subjects, eachunit containing a predetermined quantity of active component calculatedto produce the desired therapeutic effect in association with therequired pharmaceutical diluent, carrier or vehicle. The specificationfor the novel dosage forms of this invention are indicated bycharacteristics of the active component and the particular therapeuticeffect to be achieved or the limitations inherent in the art ofcompounding such an active component for therapeutic use in warm-bloodedanimals as disclosed in this specification. Examples of suitable oraldosage forms in accord with this invention are tablets, capsules, pills,powder packets, granules, wafers, cachets, teaspoonfuls, dropperfuls,ampules, vials, segregated multiples of any of the foregoing and otherforms as herein described.

The complement inhibiting activity of the compounds of this inventionhas been demonstrated by one or more of the following identified tests:(i) Test Code 026 (C1-inhibitor)--This test measures the ability ofactivated human C1 to destroy fluid phase human C2 in the presence of C4and appropriate dilutions of the test compound. An active inhibitorprotects C2 from C1 and C4; (ii) Test Code 035 (C3-C9 inhibitor)--Thistest determines the ability of the late components of human complement(C3-C9) to lyse EAC 142 in the presence of appropriate dilutions of thetest compound. An active inhibitor protects EAC 142 from lysis by humanC3-C9; (iii) Test Code 036 (C-Shunt inhibitor)--In this test humanerythrocytes rendered fragile are lysed in autologous serum via theshunt pathway activated by cobra venom factor in the presence ofappropriate dilutions of the test compound. Inhibition of the shuntpathway results in failure of lysis; (iv) Forssman VasculitisTest--Here, the well known complement dependent lesion, Forssmanvasculitis, is produced in guinea pigs by intradermal injection ofrabbit anti-Forssman antiserum. The lesion is measured in terms ofdiameter, edema and hemorrhage and the extent to which a combined indexof these is inhibited by prior intraperitoneal injection of the testcompound at 200 mg/kg is then reported, unless otherwise stated; (v)Forssman Shock Test--Lethal shock is produced in guinea pigs by an i.v.injection of anti-Forssman antiserum and the harmonic mean death time oftreated guinea pigs is compared with that of simultaneous controls; (vi)Complement Level Reduction Test--In this test, the above dosed guineapigs, or others, are bled for serum and the complement level isdetermined in undiluted serum by the capillary tube method of U.S. Pat.No. 3,876,376 and compared to undosed control guinea pigs; and (vii) Cap50 Test--Here, appropriate amounts of the test compound are added to apool of guinea pig serum in vitro, after which the undiluted serumcapillary tube assay referred to above is run. The concentration ofcompound inhibiting 50% is reported.

The results appear in Table I together with results of tests code 026,035, 036 and Cap 50, showing that the compounds of the invention possesssignificant activity compared to Suramin.

                                      TABLE I                                     __________________________________________________________________________    Biological Activities                                                                             Cl  C-Late                                                                            Shunt Inhi-                                                           026*                                                                              035*                                                                              bition 036*                                        Compound           Wells                                                                             Wells                                                                             Wells Cap 50*                                     __________________________________________________________________________    Suramin             +4**                                                                              +2**                                                                              --     361                                        8,8'-[Ureylenebis[(m-phenylenesulfonyl-                                       imino) (4-methyl-3,1-phenylenesulfonyl)-                                                          +3  +1  N     >500                                        imino]]di-1,3,5-naphthalenetrisulfonic                                        acid, hexasodium salt                                                         8,8'-[Ureylenebis[[(1,3-phenylenesul-                                         fonylimino)-1,3-phenylenesulfonyl]-                                                               +4  +5  +1**  >500                                        imino]]di-1,3,6-naphthalenetrisulfonic                                        acid, hexasodium salt                                                         8,8'-[Ureylenebis [[[(6-methyl-3,1-phen-                                      ylene)sulfonyl]imino]-[[ (6-methyl-3,1-                                       phenylene)sulfonyl]imino]]]di-1,3,6-                                                              +3  +1  N     >500                                        naphthalenetrisulfonic acid, hexa-                                            sodium salt                                                                   __________________________________________________________________________     *Code designation for tests employed as referred herein.                      **Activity in wells a serial dilution assay. Higher well number indicates     higher activity. The serial dilutions are twofold.                            N = Negative (no activity)                                               

DETAILED DESCRIPTION OF THE INVENTION EXAMPLE 18-(3-Metanilamido-p-toluenesulfonamido)-1,3,5-naphthalenetrisulfonicacid, trisodium salt

A mixture of 25.0 g of p-toluenesulfonic acid and 95.0 ml ofconcentrated nitric acid is heated on a steam bath for 30 minutes. Thesolution is poured into 250 ml of water and evaporated at reducedpressure. A small quantity of water is added and the mixture isevaporated again. This step is repeated two additional times to removeall of the nitric acid. The mixture is neutralized with a saturatedsolution of sodium carbonate and evaporated affording a yellow solid.The solid is slurried with absolute ethanol, collected by filtration andwashed twice with both ethanol and ether to give 10.3 g of product. Thefiltrate is allowed to stand and the solid formed is collected andwashed as above to provide 10.0 g of additional product and a total of20.3 g of 3-nitro-p-toluenesulfonic acid, sodium salt.

A mixture of 20.0 g of the above compound, 250 ml of thionyl chlorideand 20.0 ml of dimethylformamide is refluxed for 16 hours. The excessthionyl chloride is removed by distillation, then the mixture is cooledand ether is added. The mixture is evaporated and the crude product isdistilled under vacuum. The fraction recovered at a pressure of 1.0-1.5mm of mercury and a boiling point of 152°-154° C. provides 11.5 g of3-nitro-p-toluenesulfonyl chloride.

To a warm solution of 106.4 g of (80.5%)8-amino-1,3,5-naphthalenetrisulfonic acid in 100 ml of water and 45.0 mlof 5 N sodium hydroxide is slowly added 500 ml of absolute ethanol withvigorous stirring for 30 minutes. The mixture is cooled to roomtemperature and filtered. The precipitate is washed with 80% aqueousethanol, ethanol and ether and dried in vacuo at 110° C. for 16 hours togive 103.7 g of 8-amino-1,3,5-naphthalenetrisulfonic acid, trisodiumsalt.

An 8.0 g portion of 3-nitro-p-toluenesulfonyl chloride is added to astirred solution of 7.2 g of 8-amino-1,3,5-naphthalenetrisulfonic acidtrisodium salt, and 1.7 g of anhydrous sodium carbonate in 30.0 ml ofwater with separation of an oil. The mixture is stirred for 16 hours andthen is evaporated. The residue is dissolved in water and absoluteethanol is added to provide a precipitate. The product is collected andwashed with ethanol and ether. The filtrate is evaporated and theresidue is precipitated as above to provide additional product. A thirdcrop is obtained from the final filtrate to provide a total of 8.6 g of8-(3-nitro-p-toluenesulfonamido)-1,3,5-naphthalenetrisulfonic acid,trisodium salt.

A mixture of 8.0 g of 8-(3-nitro-p-toluenesulfonamido)-1,3,5-naphthalenetrisulfonic acid trisodium salt, 160 ml of water and 800 mg of 10%palladium on carbon catalyst is hydrogenated on a Parr shaker until noadditional hydrogen is absorbed. The resulting mixture is filteredthrough diatomaceous earth and the filtrate is evaporated. The residueis dissolved in hot water and filtered. The filtrate is triturated withethanol until cloudiness persists, then is allowed to stand at roomtemperature for 16 hours. The mixture is filtered and the filtrateevaporated to afford a gummy material which is dissolved in a smallamount of water and triturated with ethanol. An additional 200 ml ofethanol is added and the mixture is stirred for one hour to provide asolid. The solid is collected and washed with ethanol and ether to yield4.6 g of 8-(3-amino-p-toluenesulfonamido)-1,3,5-naphthalenetrisulfonicacid, trisodium salt as a light tan powder.

To a stirred solution of 2.0 g of8-(3-amino-p-toluenesulfonamido)-1,3,5-naphthalenetrisulfonic acidtrisodium salt and 342 mg of anhydrous sodium carbonate in 10 ml ofwater is added 1.5 g of m-nitrobenzenesulfonyl chloride. The mixture isstirred for 16 hours. An additional 170 mg of anhydrous sodum carbonateis added followed by 750 mg of m-nitrobenzenesulfonyl chloride and themixture is stirred for an additional 16 hours. The reaction mixture isevaporated and the residue dissolved in hot water. A product isprecipitated on the addition of absolute ethanol. The product iscollected, washed with ethanol and ether and dried. Additional productis collected from the filtrate and is washed and dried as above. Theabove fractions of product are combined and dissolved in water, then 2.5ml of 5 N sodium hydroxide is added and the mixture is stirred for 30minutes. The mixture is acidified with glacial acetic acid andevaporated. The residue is dissolved in hot water and precipitated withethanol. The product is collected and washed twice with ethanol andether and dried to yield 700 mg of8-(3-m-nitrobenzenesulfonamido-p-toluenesulfonamido)-1,3,5-naphthalenetrisulfonicacid, trisodium salt.

A mixture of 550 mg of8-(3-m-nitrobenzenesulfonamido-p-toluenesulfonamido)-1,3,5-naphthalenetrisulfonicacid, trisodium salt, 30.0 ml of water and 65.0 mg of 10% palladium oncarbon catalyst is hydrogenated on a Parr shaker for 1.75 hours. Theresulting mixture is filtered through diatomaceous earth and thefiltrate is evaporated. The crude material is dissolved in a minimum ofhot water and triturated with ethanol. The resulting solid (A) iscollected by filtration and washed with ethanol and ether. The filtrateis evaporated and the residue dissolved in water. Ethanol is added andthe solution is evaporated to yield a white powder (B). Fractions (A)and (B) are combined and dried to yield 500 mg of the desired product.

EXAMPLE 2 8,8'-[Ureylenebis[(m-phenylenesulfonylimino)(4-methyl-3,1-phenylenesulfonyl)imino]]di-1,3,5-naphthalenetrisulfonicacid, hexasodium salt

Phosgene gas is bubbled into a solution of 400 mg of the product ofExample 1, 20.0 ml of water and 55.0 mg of anhydrous sodium carbonateuntil the reaction mixture is acidic to Congo Red indicator. Thesolution is neutralized with sodium carbonate then an additional 55.0 mgof sodium carbonate is added and phosgenation is repeated until thereaction mixture is acidic. The solution is neutralized with sodiumcarbonate and the excess sodium carbonate is decomposed with aceticacid. The reaction mixture is evaporated and the residue is dissolved inhot water and filtered. The filtrate is triturated with ethanol andallowed to cool. The precipitate formed is collected and washed withethanol and ether then is re-precipitated from water and ethanol andcollected and washed as above. The product is dried to yield 336 mg ofthe desired product as a tan powder.

EXAMPLE 38-(3-Metanilamido-p-toluenesulfonamido)-1,3,6-naphthalenetrisulfonicacid, trisodium salt

Following the procedure of Example 1, employing8-amino-1,3,6-naphthalenetrisulfonic acid provides the product of theExample.

EXAMPLE 4 8,8'-[Ureylenebis[(m-phenylenesulfonfylimino)(4-methyl-3,1-phenylsulfonyl)imino]]di-1,3,6-naphthalenetrisulfonicacid, hexasodium salt

Following the procedure of Example 2, phosgenation of the product ofExample 3 provides the product of the Example.

EXAMPLE 5 8-[N³-(m-Aminophenylsulfonyl)metanilamido]-1,3,6-naphthalenetrisulfonic acid,trisodium salt

To a stirred solution of 21.9 g of 8-amino-1,3,6-naphthalenetrisulfonicacid, trisodium salt and 11.4 g of anhydrous sodium carbonate in 280 mlof water is added 24.0 g of m-nitrobenzenesulfonyl chloride. The mixtureis stirred at room temperature for 16 hours, then an additional 1.0 g ofsodium carbonate and 2.0 g of m-nitrobenzenesulfonyl chloride are addedand stirring is continued for 3 hours longer. The mixture is evaporatedand the residue is dissolved in 200 ml of water. A copious amount ofabsolute ethanol is added and the solid formed is collected and washedwith ethanol and ether, then is dried to yield 26.1 g of8-m-nitrobenzenesulfonamido-1,3,6-naphthalenetrisulfonic acid, trisodiumsalt.

A mixture of 26.1 g of8-m-nitrobenzenesulfonamido-1,3,6-naphthalenetrisulfonic acid, trisodiumsalt, 175 ml of water and 2.09 g of palladium on carbon catalyst ishydrogenated in a Parr shaker until no additional hydrogen is absorbed.The resulting mixture is filtered through diatomaceous earth and thefiltrate is evaporated. The residue is dissolved in 60.0 ml of waterand, with stirring, 400 ml of absolute ethanol is added to precipitate asolid. The mixture is allowed to stir for 2 hours, then is filtered. Theproduct is washed with absolute ethanol and ether to give 25.3 g of8-metanilamido-1,3,6-naphthalenetrisulfonic acid, trisodium salt.

To a stirred solution of 11.26 g of8-metanilamido-1,3,6-naphthalenetrisulfonic acid, trisodium salt and4.72 g of anhydrous sodium carbonate in 200 ml of water is added 10.0 gof m-nitrobenzenesulfonyl chloride. The mixture is stirred for 18 hoursand is filtered. A copious amount of absolute ethanol is added to thefiltrate, with stirring, to provide a precipitate. The mixture isstirred for one hour, then the solid is separated and washed withabsolute ethanol and ether to yield 8.9 g of 8-[N³-(m-nitrophenylsulfonyl)metanilamido]-1,3,6-naphthalenetrisulfonic acid,trisodium salt.

A mixture of 8.9 g of 8-[N³-(m-nitrophenylsulfonyl)metanilamido]-1,3,6-naphthalenetrisulfonic acid,trisodium salt, 90.0 ml of water and 1.0 g of 10% palladium on carboncatalyst is hydrogenated as previously described. The resulting mixtureis filtered through diatomaceous earth and the filtrate is evaporated.The residue is dissolved in 25.0 ml of water, then absolute ethanol isadded to precipitate the product. The precipitate is collected, iswashed with ethanol and ether and dried to yield 6.1 g of the desiredproduct.

EXAMPLE 68,8'-[Ureylenebis[[(1,3-phenylenesulfonylimino)-1,3-phenylenesulfonyl]imino]]di-1,3,6-naphthalenetrisulfonicacid, hexasodium salt

Phosgene is bubbled into a solution of 3.95 g of the product of Example5 and 2.6 ml of pyridine in 35.0 ml of water until acidic to Congo Redindicator paper. An additional 1.5 ml of pyridine is added and phosgeneis bubbled in again until acidic. The mixture is neutralized withpyridine and is poured into 450 ml of stirred absolute ethanol toprovide a gum. The supernatant is decanted, the gum is triturated withadditional absolute ethanol to yield a solid (A, (1.8 g) which iscollected and washed with ethanol and ether. The supernatant above isevaporated. The residue is triturated with a copious amount of absoluteethanol and filtered. The solid (B) is washed with ethanol and ether toprovide 2.5 g of material.

Fractions (A) and (B) above are combined and dissolved in 25.0 ml ofwater. The solution is basified to pH 8-9 with 5 N sodium hydroxide thenis neutralized with acetic acid. The solution is added dropwise to 400ml of stirred absolute ethanol to yield a precipitate. Stirring iscontinued for one hour, then the precipitate is separated, washed withabsolute ethanol and ether and dried to yield 2.0 g of the desiredproduct.

EXAMPLE 7 8-[N³-(m-Aminophenylsulfonyl)metanilamido]-1,3,5-naphthalenetrisulfonic acid,trisodium salt

Following the procedure of Example 5, employing8-amino-1,3,5-naphthalenetrisulfonic acid, trisodium salt provides theproduct of the Example.

EXAMPLE 88,8'-[Ureylenebis[[(1,3-phenylenesulfonylimino)-1,3-phenylenesulfonyl]imino]]di-1,3,5-naphthalenetrisulfonicacid, hexasodium salt

Phosgenation (according to procedure of Example 6) of the product ofExample 7 provides the product of the Example.

EXAMPLE 98-[5-(5-Amino-o-toluenesulfonamido)-o-toluenesulfonamido]-1,3,6-naphthalenetrisulfonicacid, trisodium salt

To a boiling solution of 100 g of 5-nitro-o-toluenesulfonic acid in 110ml of water is added a solution of 53.6 g of sodium chloride in 150 mlof boiling water. The reaction mixture solidifies and is heated toboiling with the addition of sufficient water to provide solution. Thensome of the water is boiled off and the mixture is allowed to stand for16 hours. The solid formed is collected and dried to yield 92.5 g of5-nitro-o-toluenesulfonic acid sodium salt.

A mixture of 50.0 g of 5-nitro-o-toluenesulfonic acid sodium salt, 125ml of thionyl chloride and 1.3 ml of dimethylformamide is stirred andrefluxed for 3 hours. The excess thionyl chloride is distilled off andthe residue is reevaporated twice with ether. The residue is extractedwith ether and methylene chloride. The extracts are evaporated and theresidue is dissolved in ether and filtered. The filtrate is concentratedwhile adding petroleum ether, then is placed in an ice box for 16 hours.The solid formed is collected and dried to yield 33.4 g of2-methyl-5-nitrobenzenesulfonyl chloride.

A mixture of 17.0 g of 8-amino-1,3,6-naphthalenetrisulfonic acid,trisodium salt, 17.5 g of 2-methyl-5-nitrobenzenesulfonyl chloride and7.8 g of anhydrous sodium carbonate in 210 ml of water is stirred atroom temperature for 18 hours, then an additional 0.5 g of sodiumcarbonate and 1.0 g of 2-methyl-5-nitrobenzenesulfonyl chloride is addedand stirring is continued for 18 hours longer. The reaction mixture isevaporated and 100 ml of water is added with stirring. The mixture isfiltered and 900 ml of absolute ethanol is added to the filtrate withstirring. The mixture is stirred for 2 hours, then the precipitate iscollected, washed with ethanol and ether and dried to yield 20.8 g of8-(5-nitro-o-toluenesulfonamido)-1,3,6-naphthalenetrisulfonic acid,trisodium salt.

A mixture of 20.0 g of8-(5-nitro-o-toluenesulfonamido)-1,3,6-naphthalenetrisulfonic acid,trisodium salt, 90.0 ml of water and 2.0 g of 10% palladium on carboncatalyst is hydrogenated as described in Example 1. The reaction mixtureis filtered through diatomaceous earth and the filtrate is evaporated.The residue is dissolved in a minimum amount of water, then is addeddropwise to 800 ml of stirred absolute ethanol. The mixture is stirredfor 2 hours and allowed to stand for 48 hours. A light yellow solid iscollected, washed with ethanol and ether, then is dried to yield 15.0 gof 8-(5-amino-o-toluenesulfonamido)-1,3,6-naphthalenetrisulfonic acid,trisodium salt.

A mixture of 7.0 g of8-(5-amino-o-toluenesulfonamido)-1,3,6-naphthalenetrisulfonic acid,trisodium salt, 5.35 g of 2-methyl-5-nitrobenzenesulfonyl chloride and2.5 g of anhydrous sodium carbonate in 120 ml of water is stirred atroom temperature for 7 hours, then an additional 0.5 g of sodiumcarbonate and 1.0 g of 2-methyl-5-nitrobenzenesulfonyl chloride is addedand stirring is continued. After 16 hours, 0.5 g of sodium carbonate and1.0 g of acid chloride is added again and stirring is continued forseveral hours. The reaction mixture is filtered and the filtrate isconcentrated and precipitated crops isolated as formed. The productcrops are combined to yield 6.0 g of8-[5-(5-nitro-o-toluenesulfonamido)-o-toluenesulfonamido]-1,3,6-naphthalenetrisulfonicacid, trisodium salt.

A mixture of 6.0 g of8-[5-(5-nitro-o-toluenesulfonamido)-o-toluenesulfonamido]-1,3,6-naphthalenetrisulfonicacid, trisodium salt, 70.0 ml of water and 920 mg of 10% palladium oncarbon catalyst is hydrogenated as previously described. The resultingmixture is filtered through diatomaceous earth and the filtrate isevaporated. The residue is dissolved in 30 ml of water and added withstirring to 300 ml of absolute ethanol forming a gum. The supernatant isdecanted and the gum is triturated with ethanol to provide a solid whichis collected by filtration and washed with ethanol and ether to yield1.4 g of product. Additional product (1.5 g) is precipitated from thesupernatant above and is collected and washed as above and the filtratesabove are evaporated and triturated with ethanol to provide 1.8 g ofproduct. The above fractions are combined and dried to yield 4.15 g ofthe desired product.

EXAMPLE 108,8'-[Ureylenebis[[[(6-methyl-3,1-phenylene)sulfonyl]imino]-[[(6-methyl-3,1-phenylene)sulfonyl]imino]]]di-1,3,6-naphthalenetrisulfonicacid, hexasodium salt

Phosgene gas is bubbled through a solution of 4.0 g of the product ofExample 9 and 2.54 ml of pyridine in 35.0 ml of water with vigorousstirring until the solution becomes acidic to Congo Red indicator paper.An additional 1.3 ml of pyridine is added and the solution isphosgenated again until acidic to Congo Red indicator. The mixture isneutralized with pyridine and poured into 650 ml of absolute ethanolwith stirring. Stirring is continued for 30 minutes and the precipitateis collected and washed with ethanol and ether. The material is dried,then is dissolved in 20 ml of water. The solution is made alkaline (pH8-9) with 5 N sodium hydroxide, then neutralized with glacial aceticacid and added to 400 ml of absolute ethanol with stirring. The solutionis concentrated to provide a precipitate. The solid is separated thenwashed with ethanol and ether and dried to yield 1.0 g of the desiredproduct.

EXAMPLE 11 8-[5-(5-Amino-o-toluenesulfonamido)-o-toluenesulfonamido]-1,3,5-naphthalenetrisulfonic acid, trisodium salt

Following the procedure of Example 9, employing8-amino-1,3,5-naphthalenetrisulfonic acid, trisodium salt provides theproduct of the Example.

EXAMPLE 128,8'-[Ureylenebis[[[(6-methyl-3,1-phenylene)sulfonyl]imino]-[[(6-methyl-3,1-phenylene)sulfonyl]imino]]]di-1,3,5-naphthalenetrisulfonicacid, hexasodium salt

Following the phosgenation procedure of Example 10, the amine of Example11 is converted into the product of the Example.

EXAMPLE 138-[3-(3-Amino-p-toluamido)-p-toluenesulfonamido]-1,3,5-naphthalenetrisulfonicacid, trisodium salt

Following the procedure of Example 1, reaction of8-(3-amino-p-toluenesulfonamido)-1,3,5-naphthalenetrisulfonic acid,trisodium salt with 3-nitro-p-toluoyl chloride followed by reductionwith palladium on carbon catalyst provides the product of the Example.

EXAMPLE 148,8'-[Ureylenebis[[[(4-methyl-3,1-phenylenecarbonyl)imino](4-methyl-3,1-phenylenesulfonyl)]imino]]di-1,3,5-naphthalenetrisulfonicacid, hexasodium salt

Following the procedure of Example 2, the amino product of Example 13 istreated with phosgene to produce the ureylene, the product of theExample.

EXAMPLE 158-[3-(3-Amino-p-toluamido)-p-toluenesulfonamido]-1,3,6-naphthalenetrisulfonicacid, trisodium salt

Following the procedure of Example 1 with8-amino-1,3,6-naphthalenetrisulfonic acid, trisodium salt provides8-(3-amino-p-toluenesulfonamido)-1,3,6-naphthalenetrisulfonic acid,trisodium salt. Reaction of the latter with 3-nitro-p-toluoyl chloridefollowed by reduction with palladium on carbon catalyst provides theproduct of the Example.

EXAMPLE 168,8'-[Ureylenebis[[[(4-methyl-3,1-phenylenecarbonyl)imino](4-methyl-3,1-phenylenesulfonyl)]imino]]di-1,3,6-naphthalenetrisulfonicacid, hexasodium salt

Following the procedure of Example 2, the amino product of Example 15 istreated with phosgene to produce the ureylene, the product of theExample.

EXAMPLE 178-[3-(3-Aminobenzamido)-p-toluenesulfonamido]-1,3,6-naphthalenetrisulfonicacid, trisodium salt

Following the procedure of Example 1, reaction of8-(3-amino-p-toluenesulfonamido)-1,3,6-naphthalenetrisulfonic acid,trisodium salt (Example 14) with 3-nitrobenzoyl chloride followed byreduction with palladium on carbon catalyst provides the product of theExample.

EXAMPLE 188,8'-[Ureylenebis[[(3,1-phenylenecarbonylimino)(4-methyl-3,1-phenylenesulfonyl)]imino]]di-1,3,6-naphthalenetrisulfonicacid, hexasodium salt

Following the procedure of Example 2, the amino product of Example 17 istreated with phosgene to produce the ureylene, the product of theExample.

EXAMPLE 198-[3-(3-Aminobenzamido)-p-toluenesulfonamido]-1,3,5-naphthalenetrisulfonicacid, trisodium salt

Following the procedure of Example 1, reaction of8-(3-amino-p-toluenesulfonamido)-1,3,5-naphthalenetrisulfonic acid,trisodium salt with 3-nitrobenzoyl chloride followed by reduction withpalladium on carbon catalyst provides the product of the Example.

EXAMPLE 208,8'-[Ureylenebis[[(3,1-phenylenecarbonylimino)(4-methyl-3,1-phenylenesulfonyl)]imino]]di-1,3,5-naphthalenetrisulfonicacid, hexasodium salt

Following the procedure of Example 2, the amino product of Example 19 istreated with phosgene to produce the ureylene, the product of theExample.

EXAMPLE 218-[5-(5-Amino-o-toluamido)-o-toluenesulfonamido]-1,3,5-naphthalenetrisulfonicacid, trisodium salt

Following the procedure of Example 1, reaction of8-amino-1,3,5-naphthalenetrisulfonic acid with 5-nitro-o-toluenesulfonylchloride provides8-(5-nitro-o-toluenesulfonamido)-1,3,5-naphthalenetrisulfonic acid,trisodium salt, followed by reduction with palladium on carbon catalystprovides 8-(5-amino-o-toluenesulfonamido)-1,3,5-naphthalenetrisulfonicacid, trisodium salt.

The preceding product is then reacted with 5-nitro-o-toluoyl chloride toyield8-[5-(5-amino-o-toluamido)-o-toluenesulfonamido]-1,3,5-naphthalenetrisulfonicacid, trisodium salt.

EXAMPLE 228,8'-[Ureylenebis[[[(6-methyl-3,1-phenylenecarbonyl)imino](6-methyl-3,1-phenylenesulfonyl)]imino]]di-1,3,5-naphthalenetrisulfonicacid, hexasodium salt

Following the procedure of Example 2, the amino product of Example 21 istreated with phosgene to produce the ureylene, the product of theExample.

EXAMPLE 238-[5-(5-Amino-o-toluamido)-o-toluenesulfonamido]-1,3,6-naphthalenetrisulfonicacid, trisodium salt

Following the procedure of Example 1, reaction of8-amino-1,3,6-napthalenetrisulfonic acid, with 5-nitro-o-toluenesulfonylchloride provides8-(5-nitro-o-toluenesulfonamido)-1,3,5-naphthalenetrisulfonic acid,trisodium salt, followed by reduction with palladium on carbon catalystprovides 8-(5-amino-o-toluenesulfonamido)-1,3,5-naphthalenetrisulfonicacid, trisodium salt.

The preceding product is then reacted with 5-nitro-o-toluoyl chloride toyield8-[5-(5-amino-o-toluamido)-o-toluenesulfonamido]-1,3,5-naphthalenetrisulfonicacid, trisodium salt.

EXAMPLE 248,8'-([Ureylenebis[[[(6-methyl-3,1-phenylenecarbonyl)imino](6-methyl-3,1-phenylenesulfonyl)]imino]]di-1,3,6-naphthalenetrisulfonicacid, hexasodium salt

Following the procedure of Example 2, the amino product of Example 23 istreated with phosgene to produce the ureylene, the product of theExample.

EXAMPLE 258-[N-(m-Aminobenzoyl)metanilamido]-1,3,5-naphthalenetrisulfonic acid,trisodium salt

Following the procedure of Example 1, reaction of8-amino-1,3,5-naphthalenetrisulfonic acid, trisodium salt with3-nitrobenzenesulfonyl chloride provides, after reduction,8-(metanilamido)-1,3,5-naphthalenetrisulfonic acid, trisodium salt.Treatment with m-nitrobenzoyl chloride, reducing the product therefromprovides the product of the Example.

EXAMPLE 268,8'-[Ureylenebis[[(3,1-phenylenecarbonylimino)-3,1-phenylenesulfonyl]imino]]di-1,3,5-naphthalene-trisulfonicacid, hexasodium salt

Following the procedure of Example 2, treatment of the product ofExample 25 with phosgene generates the ureylene, the product of theExample.

EXAMPLE 278-[N-(m-Aminobenzoyl)-metanilamido]-1,3,6-naphthalenetrisulfonic acid,trisodium salt

Following the procedure of Example 1, reaction of8-amino-1,3,6-naphthalenetrisulfonic acid, trisodium salt with3-nitrobenzenesulfonyl chloride provides, after reduction,8-(metanilamido)-1,3,6-naphthalenetrisulfonic acid, trisodium salt.Treatment with m-nitrobenzoyl chloride, reducing the product therefromprovides the product of the Example.

EXAMPLE 288,8'-[Ureylenebis[[(3,1-phenylenecarbonylimino)-3,1-phenylenesulfonyl]imino]]di-1,3,6-naphthalenetrisulfonicacid, hexasodium salt

Following the procedure of Example 2, treatment of the product ofExample 27 with phosgene generates the ureylene, the product of theExample.

EXAMPLE 298-[3-(5-Amino-2,4-dimethylbenzamido)-p-toluene-sulfonamido]-1,3,5-naphthalenetrisulfonicacid, trisodium salt

Following the procedure of Example 1, reaction of8-(3-amino-p-toluenesulfonamido)-1,3,5-naphthalenetrisulfonic acid,trisodium salt with 5-nitro-2,4-dimethylbenzoyl chloride generates8-[3-(5-nitro-2,4-dimethylbenzamido)-p-toluenesulfonamido]-1,3,5-naphthalenetrisulfonicacid, trisodium salt. Reduction with palladium on carbon catalyst givesthe product of the Example.

EXAMPLE 308,8'-[Ureylenebis[[[(4,6-dimethyl-3,1-phenylenecarbonyl)imino](4-methyl-3,1-phenylenesulfonyl)]imino]]di-1,3,5-naphthalenetrisulfonicacid, hexasodium salt

Following the procedure of Example 2, phosgenation of the product fromExample 29 provides the proudct of the Example.

EXAMPLE 318-[3-(5-Amino-2,4-dimethylbenzamido)-p-toluenesulfonamido]-1,3,6-naphthalenetrisulfonicacid, trisodium salt

Following the procedure of Example 1, reaction of8-(3-amino-p-toluenesulfonamido)-1,3,6-naphthalenetrisulfonic acid,trisodium salt with 5-nitro-2,4-dimethylbenzoyl chloride generates8-[3-(5-nitro-2,4-dimethylbenzamido)-p-toluenesulfonamido]-1,3,6-naphthalenetrisulfonicacid, trisodium salt. Reduction with palladium on carbon catalyst givesthe product of the Example.

EXAMPLE 328,8'-[Ureylenebis[[[(4,6-dimethyl-3,1-phenylene-carbonyl)imino](4-methyl-3,1-phenylenesulfonyl)]imino]]di-1,3,6-naphthalenetrisulfonicacid, hexasodium salt

Following the procedure of Example 2, phosgenation of the product fromExample 31 provides the product of the Example.

EXAMPLE 33 Preparation of Compressed Tablet

    ______________________________________                                        Ingredient           mg/Tablet                                                ______________________________________                                        Active Compound       0.5-500                                                 Dibasic Calcium Phosphate N.F.                                                                     qs                                                       Starch USP           40                                                       Modified Starch      10                                                       Magnesium Stearate USP                                                                             1-5                                                      ______________________________________                                    

EXAMPLE 34 Preparation of Compressed Tablet--Sustained Action T1Ingredient? mg/Tablet? Active Compound as Aluminum 0.5-500 (as acidLake*, Micronized equivalent) Dibasic Calcium Phosphate N.F. qs AlginicAcid 20 EXAMPLE 35 Preparation of Hard Shell Capsule

    ______________________________________                                        Ingredient        mg/Capsule                                                  ______________________________________                                        Active Compound   0.5-500                                                     Lactose, Spray Dried                                                                            qs                                                          Magnesium Stearate                                                                              1-10                                                        ______________________________________                                    

EXAMPLE 36 Preparation of Oral Liquid (Syrup)

    ______________________________________                                        Ingredient        % W/V                                                       ______________________________________                                        Active Compound   0.05-5                                                      Liquid Sugar      75.0                                                        Methyl Paraben USP                                                                              0.18                                                        Propyl Paraben USP                                                                              0.02                                                        Flavoring Agent   qs                                                          Purified Water qs ad                                                                            100.0                                                       ______________________________________                                    

EXAMPLE 37 Preparation of Oral Liquid (Elixir)

    ______________________________________                                        Ingredient        % W/V                                                       ______________________________________                                        Active Compound   0.05-5                                                      Alcohol USP       12.5                                                        Glycerin USP      45.0                                                        Syrup USP         20.0                                                        Flavoring Agent   qs                                                          Purified Water qs ad                                                                            100.0                                                       ______________________________________                                    

EXAMPLE 38 Preparation of Oral Suspension (Syrup)

    ______________________________________                                        Ingredient          % W/V                                                     ______________________________________                                        Active Compound as Aluminum                                                                       0.05-5                                                    Lake, Micronized    (acid equivalent)                                         Polysorbate 80 USP  0.1                                                       Magnesium Aluminum Silicate,                                                  Colloidal           0.3                                                       Flavoring Agent     qs                                                        Methyl Paraben USP  0.18                                                      Propyl Paraben USP  0.02                                                      Liquid Sugar        75.0                                                      Purified Water qs ad                                                                              100.0                                                     ______________________________________                                    

EXAMPLE 39 Preparation of Injectable Solution

    ______________________________________                                        Ingredient        % W/V                                                       ______________________________________                                        Active Compound   0.05-5                                                      Benzyl Alcohol N.F.                                                                             0.9                                                         Water for Injection                                                                             100.0                                                       ______________________________________                                    

EXAMPLE 40 Preparation of Injectable Oil

    ______________________________________                                        Ingredient        % W/V                                                       ______________________________________                                        Active Compound   0.05-5                                                      Benzyl Alcohol    1.5                                                         Sesame Oil qs ad  100.0                                                       ______________________________________                                    

EXAMPLE 41 Preparation of Intra-Articular Product

    ______________________________________                                        Ingredient             Amount                                                 ______________________________________                                        Active Compound        2-20 mg                                                NaCl (physiological saline)                                                                          0.9%                                                   Benzyl Alcohol         0.9%                                                   Sodium Carboxymethylcellulose                                                                        1-5%                                                   pH adjusted to 5.0-7.5                                                        Water for Injection qs ad                                                                            100%                                                   ______________________________________                                    

EXAMPLE 42 Preparation of Injectable Depo Suspension

    ______________________________________                                        Ingredient          % W/V                                                     ______________________________________                                        Active Compound     0.05-5                                                                        (acid equivalent)                                         Polysorbate 80 USP  0.2                                                       Polyethylene Glycol 4000 USP                                                                      3.0                                                       Sodium Chloride USP 0.8                                                       Benzyl Alcohol N.F. 0.9                                                       HCl to pH 6-8       qs                                                        Water for Injection qs ad                                                                         100.0                                                     ______________________________________                                    

EXAMPLE 43 Preparation of Dental Paste

    ______________________________________                                        Ingredient          % W/W                                                     ______________________________________                                        Active Compound     0.05-5                                                    Zinc Oxide          15                                                        Polyethylene Glycol 4000 USP                                                                      50                                                        Distilled Water qs  100                                                       ______________________________________                                    

EXAMPLE 44 Preparation of Dental Ointment

    ______________________________________                                        Ingredient          % W/W                                                     ______________________________________                                        Active Compound     0.05-5                                                    Petrolatum, White USP qs                                                                          100                                                       ______________________________________                                    

EXAMPLE 45 Preparation of Dental Cream

    ______________________________________                                        Ingredient          % W/W                                                     ______________________________________                                        Active Compound     0.05-5                                                    Mineral Oil         50                                                        Beeswax             15                                                        Sorbitan Monostearate                                                                             2                                                         Polyoxyethylene 20 Sorbitan                                                   Monostearate        3                                                         Methylparaben USP   0.18                                                      Propyl Paraben USP  0.02                                                      Distilled Water qs  100                                                       ______________________________________                                    

EXAMPLE 46 Preparation of Topical Cream

    ______________________________________                                        Ingredient          % W/W                                                     ______________________________________                                        Active Compound     0.05-5                                                    Sodium Lauryl Sulfate                                                                             1                                                         Propylene Glycol    12                                                        Stearyl Alcohol     25                                                        Petrolatum, White USP                                                                             25                                                        Methyl Paraben USP  0.18                                                      Propyl Paraben USP  0.02                                                      Purified Water qs   100                                                       ______________________________________                                    

EXAMPLE 47 Preparation of Topical Ointment

    ______________________________________                                        Ingredient          % W/W                                                     ______________________________________                                        Active Compound     0.05-5                                                    Cholesterol         3                                                         Stearyl Alcohol     3                                                         White Wax           8                                                         Petrolatum, White USP qs                                                                          100                                                       ______________________________________                                    

EXAMPLE 48 Preparation of Spray Lotion (non-Aerosol)

    ______________________________________                                        Ingredient          % W/W                                                     ______________________________________                                        Active Compound     0.05-5                                                    Isopropyl Myristate  20                                                       Alcohol (Denatured) qs                                                                            100                                                       ______________________________________                                    

EXAMPLE 49 Preparation of Buccal Tablet

    ______________________________________                                        Ingredient            g/Tablet                                                ______________________________________                                        Active Ingredient     0.00325                                                 6 × Sugar       0.29060                                                 Acacia                0.01453                                                 Soluble Starch        0.01453                                                 F. D. & C. Yellow No. 6 Dye                                                                         0.00049                                                 Magnesium Stearate    0.00160                                                                       0.32500                                                 ______________________________________                                    

The final tablet will weigh about 325 mg. and may be compressed intobuccal tablets in flat faced or any other tooling shape convenient forbuccal administration.

EXAMPLE 50 Preparation of Lozenge

    ______________________________________                                        Ingredient           g/Lozenge                                                ______________________________________                                        Active Ingredient    0.0140                                                   Kompact® Sugar (Sucrest Co.)                                                                   0.7138                                                   6 × Sugar      0.4802                                                   Sorbitol (USP Crystalline)                                                                         0.1038                                                   Flavor               0.0840                                                   Magnesium Stearate   0.0021                                                   Dye                  qs                                                       Stearic Acid         0.0021                                                                        1.4000                                                   ______________________________________                                    

The ingredients are compressed into 5/8" flat based lozenge tooling.Other shapes may also be utilized.

We claim:
 1. A compound of the formula: ##STR4## wherein R, R₁, R₂ andR₃ are selected from the group consisting of hydrogen and methyl; and Ais a pharmaceutically acceptable salt cation.
 2. The compound accordingto claim 1,8-[3-(3-Amino-p-toluamido)-p-toluenesulfonamido]-1,3,5-naphthalene-trisulfonicacid, trisodium salt.
 3. The compound according to claim 1,8-[3-(3-Amino-p-toluamido)-p-toluenesulfonamido]-1,3,6-naphthalenetrisulfonicacid, trisodium salt.
 4. The compound according to claim 1,8-[3-(3-Aminobenzamido)-p-toluenesulfonamido[-1,3,6-naphthalenetrisulfonicacid, trisodium salt.
 5. The compound according to claim 1,8-[3-(3-Aminobenzamido)-p-toluenesulfonamido]-1,3,5-naphthalenetrisulfonicacid, trisodium salt.
 6. The compound according to claim 1,8-[5-(5-Amino-o-toluamido)-o-toluenesulfonamido]-1,3,5-naphthalenetrisulfonicacid, trisodium salt.
 7. The compound according to claim 1,8-[5-(5-Amino-o-toluamido)-o-toluenesulfonamido]-1,3,6-naphthalenetrisulfonicacid, trisodium salt.
 8. The compound according to claim 1,8-[N-(m-Aminobenzoyl)metanilamido]-1,3,5-naphthalenetrisulfonic acid,trisodium salt.
 9. The compound according to claim 1,8-[N-(m-Aminobenzoyl)metanilamido]-1,3,6-naphthalenetrisulfonic acid,trisodium salt.
 10. The compound according to claim 1,8-[3-(5-Amino-2,4-dimethylbenzamido)-p-toluenesulfonamido]-1,3,5-naphthalenetrisulfonicacid, trisodium salt.
 11. The compound according to claim 1,8-[3-(5-Amino-2,4-dimethylbenzamido)-p-toluenesulfonamido]-1,3,6-naphthalenetrisulfonicacid, trisodium salt.
 12. A compound of the formula: ##STR5## wherein R,R₁, R₂ and R₃ are selected from the group consisting of hydrogen andmethyl; and A is a pharmaceutically acceptable salt cation.
 13. Thecompound according to claim 12,8-(3-Metanilamido-p-toluenesulfonamido)-1,3,5-naphthalenetrisulfonicacid, trisodium salt.
 14. The compound according to claim 12,8-(3-Metanilamido-p-toluenesulfonamido)-1,3,6-naphthalenetrisulfonicacid, trisodium salt.
 15. The compound according to claim 12, 8-[N³-(m-Aminophenylsulfonyl)metanilamido]-1,3,6-naphthalenetrisulfonic acid,trisodium salt.
 16. The compound according to claim 12, 8-[N³-(m-Aminophenylsulfonyl)metanilamido]-1,3,5-naphthalenetrisulfonic acid,trisodium salt.
 17. The compound according to claim 12,8-[5-(5-Amino-o-toluenesulfonamido)-o-toluenesulfonamido]-1,3,6-naphthalenetrisulfonicacid, trisodium salt.
 18. The compound according to claim 12,8-[5-(5-Amino-o-toluenesulfonamido)-o-toluenesulfonamido]-1,3,5-naphthalenetrisulfonicacid, trisodium salt.